Collagen density regulates the activity of tumor-infiltrating T cells

Dorota E Kuczek, Anne Mette H Larsen, Marie-Louise Thorseth, Marco Carretta, Adrija Kalvisa, Majken S Siersbæk, Ana Micaela C Simões, Anne Roslind, Lars H Engelholm, Elfriede Noessner, Marco Donia, Inge Marie Svane, Per Thor Straten, Lars Grøntved, Daniel H Madsen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

103 Downloads (Pure)

Abstract

BACKGROUND: Tumor progression is accompanied by dramatic remodeling of the surrounding extracellular matrix leading to the formation of a tumor-specific ECM, which is often more collagen-rich and of increased stiffness. The altered ECM of the tumor supports cancer growth and metastasis, but it is unknown if this effect involves modulation of T cell activity. To investigate if a high-density tumor-specific ECM could influence the ability of T cells to kill cancer cells, we here studied how T cells respond to 3D culture in different collagen densities.

METHODS: T cells cultured in 3D conditions surrounded by a high or low collagen density were imaged using confocal fluorescent microscopy. The effects of the different collagen densities on T cell proliferation, survival, and differentiation were examined using flow cytometry. Cancer cell proliferation in similar 3D conditions was also measured. Triple-negative breast cancer specimens were analyzed for the number of infiltrating CD8+ T cells and for the collagen density. Whole-transcriptome analyses were applied to investigate in detail the effects of collagen density on T cells. Computational analyses were used to identify transcription factors involved in the collagen density-induced gene regulation. Observed changes were confirmed by qRT-PCR analysis.

RESULTS: T cell proliferation was significantly reduced in a high-density matrix compared to a low-density matrix and prolonged culture in a high-density matrix led to a higher ratio of CD4+ to CD8+ T cells. The proliferation of cancer cells was unaffected by the surrounding collagen-density. Consistently, we observed a reduction in the number of infiltrating CD8+ T-cells in mammary tumors with high collagen-density indicating that collagen-density has a role in regulating T cell abundance in human breast cancer. Whole-transcriptome analysis of 3D-cultured T cells revealed that a high-density matrix induces downregulation of cytotoxic activity markers and upregulation of regulatory T cell markers. These transcriptional changes were predicted to involve autocrine TGF-β signaling and they were accompanied by an impaired ability of tumor-infiltrating T cells to kill autologous cancer cells.

CONCLUSIONS: Our study identifies a new immune modulatory mechanism, which could be essential for suppression of T cell activity in the tumor microenvironment.

Original languageEnglish
Article number68
JournalJournal for immunotherapy of cancer
Volume7
Issue number1
Number of pages15
ISSN2051-1426
DOIs
Publication statusPublished - 12. Mar 2019

Fingerprint

Neoplasms
Cell Proliferation
Gene Expression Profiling
Autocrine Communication
CD4-CD8 Ratio
Confocal Microscopy
Cell Differentiation
Cultured Cells
Cell Survival
Flow Cytometry
Up-Regulation
Down-Regulation
Polymerase Chain Reaction
Growth

Keywords

  • 3D culture
  • Extracellular matrix
  • Immune modulation
  • T cell activity
  • Tumor microenvironment

Cite this

Kuczek, D. E., Larsen, A. M. H., Thorseth, M-L., Carretta, M., Kalvisa, A., Siersbæk, M. S., ... Madsen, D. H. (2019). Collagen density regulates the activity of tumor-infiltrating T cells. Journal for immunotherapy of cancer, 7(1), [68]. https://doi.org/10.1186/s40425-019-0556-6
Kuczek, Dorota E ; Larsen, Anne Mette H ; Thorseth, Marie-Louise ; Carretta, Marco ; Kalvisa, Adrija ; Siersbæk, Majken S ; Simões, Ana Micaela C ; Roslind, Anne ; Engelholm, Lars H ; Noessner, Elfriede ; Donia, Marco ; Svane, Inge Marie ; Straten, Per Thor ; Grøntved, Lars ; Madsen, Daniel H. / Collagen density regulates the activity of tumor-infiltrating T cells. In: Journal for immunotherapy of cancer. 2019 ; Vol. 7, No. 1.
@article{0dce9a17329f4c6b9971e2915e3fec22,
title = "Collagen density regulates the activity of tumor-infiltrating T cells",
abstract = "BACKGROUND: Tumor progression is accompanied by dramatic remodeling of the surrounding extracellular matrix leading to the formation of a tumor-specific ECM, which is often more collagen-rich and of increased stiffness. The altered ECM of the tumor supports cancer growth and metastasis, but it is unknown if this effect involves modulation of T cell activity. To investigate if a high-density tumor-specific ECM could influence the ability of T cells to kill cancer cells, we here studied how T cells respond to 3D culture in different collagen densities.METHODS: T cells cultured in 3D conditions surrounded by a high or low collagen density were imaged using confocal fluorescent microscopy. The effects of the different collagen densities on T cell proliferation, survival, and differentiation were examined using flow cytometry. Cancer cell proliferation in similar 3D conditions was also measured. Triple-negative breast cancer specimens were analyzed for the number of infiltrating CD8+ T cells and for the collagen density. Whole-transcriptome analyses were applied to investigate in detail the effects of collagen density on T cells. Computational analyses were used to identify transcription factors involved in the collagen density-induced gene regulation. Observed changes were confirmed by qRT-PCR analysis.RESULTS: T cell proliferation was significantly reduced in a high-density matrix compared to a low-density matrix and prolonged culture in a high-density matrix led to a higher ratio of CD4+ to CD8+ T cells. The proliferation of cancer cells was unaffected by the surrounding collagen-density. Consistently, we observed a reduction in the number of infiltrating CD8+ T-cells in mammary tumors with high collagen-density indicating that collagen-density has a role in regulating T cell abundance in human breast cancer. Whole-transcriptome analysis of 3D-cultured T cells revealed that a high-density matrix induces downregulation of cytotoxic activity markers and upregulation of regulatory T cell markers. These transcriptional changes were predicted to involve autocrine TGF-β signaling and they were accompanied by an impaired ability of tumor-infiltrating T cells to kill autologous cancer cells.CONCLUSIONS: Our study identifies a new immune modulatory mechanism, which could be essential for suppression of T cell activity in the tumor microenvironment.",
keywords = "3D culture, Extracellular matrix, Immune modulation, T cell activity, Tumor microenvironment",
author = "Kuczek, {Dorota E} and Larsen, {Anne Mette H} and Marie-Louise Thorseth and Marco Carretta and Adrija Kalvisa and Siersb{\ae}k, {Majken S} and Sim{\~o}es, {Ana Micaela C} and Anne Roslind and Engelholm, {Lars H} and Elfriede Noessner and Marco Donia and Svane, {Inge Marie} and Straten, {Per Thor} and Lars Gr{\o}ntved and Madsen, {Daniel H}",
year = "2019",
month = "3",
day = "12",
doi = "10.1186/s40425-019-0556-6",
language = "English",
volume = "7",
journal = "Journal for immunotherapy of cancer",
issn = "2051-1426",
publisher = "BioMed Central",
number = "1",

}

Kuczek, DE, Larsen, AMH, Thorseth, M-L, Carretta, M, Kalvisa, A, Siersbæk, MS, Simões, AMC, Roslind, A, Engelholm, LH, Noessner, E, Donia, M, Svane, IM, Straten, PT, Grøntved, L & Madsen, DH 2019, 'Collagen density regulates the activity of tumor-infiltrating T cells', Journal for immunotherapy of cancer, vol. 7, no. 1, 68. https://doi.org/10.1186/s40425-019-0556-6

Collagen density regulates the activity of tumor-infiltrating T cells. / Kuczek, Dorota E; Larsen, Anne Mette H; Thorseth, Marie-Louise; Carretta, Marco; Kalvisa, Adrija; Siersbæk, Majken S; Simões, Ana Micaela C; Roslind, Anne; Engelholm, Lars H; Noessner, Elfriede; Donia, Marco; Svane, Inge Marie; Straten, Per Thor; Grøntved, Lars; Madsen, Daniel H.

In: Journal for immunotherapy of cancer, Vol. 7, No. 1, 68, 12.03.2019.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Collagen density regulates the activity of tumor-infiltrating T cells

AU - Kuczek, Dorota E

AU - Larsen, Anne Mette H

AU - Thorseth, Marie-Louise

AU - Carretta, Marco

AU - Kalvisa, Adrija

AU - Siersbæk, Majken S

AU - Simões, Ana Micaela C

AU - Roslind, Anne

AU - Engelholm, Lars H

AU - Noessner, Elfriede

AU - Donia, Marco

AU - Svane, Inge Marie

AU - Straten, Per Thor

AU - Grøntved, Lars

AU - Madsen, Daniel H

PY - 2019/3/12

Y1 - 2019/3/12

N2 - BACKGROUND: Tumor progression is accompanied by dramatic remodeling of the surrounding extracellular matrix leading to the formation of a tumor-specific ECM, which is often more collagen-rich and of increased stiffness. The altered ECM of the tumor supports cancer growth and metastasis, but it is unknown if this effect involves modulation of T cell activity. To investigate if a high-density tumor-specific ECM could influence the ability of T cells to kill cancer cells, we here studied how T cells respond to 3D culture in different collagen densities.METHODS: T cells cultured in 3D conditions surrounded by a high or low collagen density were imaged using confocal fluorescent microscopy. The effects of the different collagen densities on T cell proliferation, survival, and differentiation were examined using flow cytometry. Cancer cell proliferation in similar 3D conditions was also measured. Triple-negative breast cancer specimens were analyzed for the number of infiltrating CD8+ T cells and for the collagen density. Whole-transcriptome analyses were applied to investigate in detail the effects of collagen density on T cells. Computational analyses were used to identify transcription factors involved in the collagen density-induced gene regulation. Observed changes were confirmed by qRT-PCR analysis.RESULTS: T cell proliferation was significantly reduced in a high-density matrix compared to a low-density matrix and prolonged culture in a high-density matrix led to a higher ratio of CD4+ to CD8+ T cells. The proliferation of cancer cells was unaffected by the surrounding collagen-density. Consistently, we observed a reduction in the number of infiltrating CD8+ T-cells in mammary tumors with high collagen-density indicating that collagen-density has a role in regulating T cell abundance in human breast cancer. Whole-transcriptome analysis of 3D-cultured T cells revealed that a high-density matrix induces downregulation of cytotoxic activity markers and upregulation of regulatory T cell markers. These transcriptional changes were predicted to involve autocrine TGF-β signaling and they were accompanied by an impaired ability of tumor-infiltrating T cells to kill autologous cancer cells.CONCLUSIONS: Our study identifies a new immune modulatory mechanism, which could be essential for suppression of T cell activity in the tumor microenvironment.

AB - BACKGROUND: Tumor progression is accompanied by dramatic remodeling of the surrounding extracellular matrix leading to the formation of a tumor-specific ECM, which is often more collagen-rich and of increased stiffness. The altered ECM of the tumor supports cancer growth and metastasis, but it is unknown if this effect involves modulation of T cell activity. To investigate if a high-density tumor-specific ECM could influence the ability of T cells to kill cancer cells, we here studied how T cells respond to 3D culture in different collagen densities.METHODS: T cells cultured in 3D conditions surrounded by a high or low collagen density were imaged using confocal fluorescent microscopy. The effects of the different collagen densities on T cell proliferation, survival, and differentiation were examined using flow cytometry. Cancer cell proliferation in similar 3D conditions was also measured. Triple-negative breast cancer specimens were analyzed for the number of infiltrating CD8+ T cells and for the collagen density. Whole-transcriptome analyses were applied to investigate in detail the effects of collagen density on T cells. Computational analyses were used to identify transcription factors involved in the collagen density-induced gene regulation. Observed changes were confirmed by qRT-PCR analysis.RESULTS: T cell proliferation was significantly reduced in a high-density matrix compared to a low-density matrix and prolonged culture in a high-density matrix led to a higher ratio of CD4+ to CD8+ T cells. The proliferation of cancer cells was unaffected by the surrounding collagen-density. Consistently, we observed a reduction in the number of infiltrating CD8+ T-cells in mammary tumors with high collagen-density indicating that collagen-density has a role in regulating T cell abundance in human breast cancer. Whole-transcriptome analysis of 3D-cultured T cells revealed that a high-density matrix induces downregulation of cytotoxic activity markers and upregulation of regulatory T cell markers. These transcriptional changes were predicted to involve autocrine TGF-β signaling and they were accompanied by an impaired ability of tumor-infiltrating T cells to kill autologous cancer cells.CONCLUSIONS: Our study identifies a new immune modulatory mechanism, which could be essential for suppression of T cell activity in the tumor microenvironment.

KW - 3D culture

KW - Extracellular matrix

KW - Immune modulation

KW - T cell activity

KW - Tumor microenvironment

U2 - 10.1186/s40425-019-0556-6

DO - 10.1186/s40425-019-0556-6

M3 - Journal article

C2 - 30867051

VL - 7

JO - Journal for immunotherapy of cancer

JF - Journal for immunotherapy of cancer

SN - 2051-1426

IS - 1

M1 - 68

ER -